J Arthropod-Borne Dis, September 2020, 14(3): 293–301 S Sukumaran and R Maheswaran: Larvicidal Activity of … 293 http://jad.tums.ac.ir Published Online: September 30, 2020 Short Communication Larvicidal Activity of Elytraria acaulis against Culex quinquefasciatus and Aedes aegypti (Diptera: Culicidae) Soorya Sukumaran, *Rajan Maheswaran Department of Zoology, Entomology Laboratory, School of Life Sciences, Periyar University, Salem, Tamil Nadu, India *Corresponding author: Dr Rajan Maheswaran, E-mail: mahes1380@gmail.com (Received 05 Feb 2016; accepted 18 Jul 2020) Abstract Background: Mosquitoes are blood sucking arthropods and serve as vectors of many diseases causing serious health problems to human beings. Culex quinquefasciatus and Aedes aegypti were responsible for Filariasis and Dengue. Syn- thetic pesticides were effective against mosquitoes as well as main sources of environmental pollution and most of them are immunosuppressant. Botanicals were widely used as insecticides, growth disruptors, repellents, etc. The aim of this research was to determine larvicidal properties of powdered leaf, Elytraria acaulis against late third or early fourth in- star larvae of Cx. quinquefasciatus and Ae. aegypti. Methods: Larvae of Cx. quinquefasciatus and Ae. aegypti were tested at various concentrations of 100, 120, 140, 160, 180 and 200mg/100ml and mortality was recorded after 24h. The LC50 values of the E. acaulis leaf powder were calcu- lated by Probit analysis. Results: The plant powder exhibited strong larvicidal activity against Cx. quinquefasciatus with LC50 value of 116.07mg/100ml against Ae. aegypti 124.25mg/100ml respectively. The result indicated that the plant powder of E. acaulis showed potential larvicidal activity against Cx. quinquefasciatus and Ae. aegypti. Conclusion: The overall findings of the present investigation suggested that the E. acaulis highly effective against Cx. quinquefasciatus and Ae. aegypti larvae. Elytraria acaulis may be used as an alternative to synthetic chemical pesticides for control of vectors to reduce vector borne diseases and did not harm to total environment. Keywords: Elytraria acaulis; Larvicidal activity; Culex quinquefasciatus; Aedes aegypti Introduction Man suffers extensively due to the nuisance of vector mosquito population in public health manner. Mosquitoes directly transmit diseases such as filarial, malaria and dengue fever. Mos- quitoes are blood sucking insects and serve as vectors for spreading human diseases and there- fore, they continue to pose a serious health prob- lem throughout the world. These are not only the most important vector for the transmission of diseases (1) but also an important pest to hu- mans, causing allergic responses that include local skin reaction and systemic reaction such angio-edema and urticaria (2). Culex quinque- fasciatus, a vector of lymphatic filariasis, is widely distributed in tropical zones with around 120 million people infected worldwide and 44 million people having common chronic mani- festation (3). Despite its debilitating effects, lym- phatic filariasis is given a very low control pri- ority (4). In most of its range the females in- tensely anthropophilic, fed actively only at night and it causes nuisance (5) and are vectors of Japanese encephalitis, West Nile virus St. Louis encephalitis and avian malaria. Aedes aegypti, the principal vector of dengue, chikungunya, Zika and yellow fever viruses, is an anthro- pophilic species adapted to urban environments, particularly to housing (6). Dengue Hemorrhag- ic Fever (DHF) and Chikungunya are the ma- jor mosquito-borne diseases in India. The first dengue hemorrhagic fever was reported in Thai- land and Philippines in 1950s. Dengue infec- tions are reported throughout the world including India, where the first dengue outbreak was re- Copyright © 2020 Iranian Scientific Society of Biology & Control of Diseases Vectors, and Tehran University of Medical Sciences. Published by Tehran University of Medical Sciences. This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International license (https://creativecommons.org/licenses/by-nc/4.0/). Non-commercial uses of the work are permitted, provided the original work is properly cited. http://jad.tums.ac.ir/ mailto:mahes1380@gmail.com https://creativecommons.org/licenses/by-nc/4.0/ J Arthropod-Borne Dis, September 2020, 14(3): 293–301 S Sukumaran and R Maheswaran: Larvicidal Activity of … 294 http://jad.tums.ac.ir Published Online: September 30, 2020 ported in Delhi in 1988 (7). Now the dengue infection has been reported from all over the country (8-13) with major outbreaks reported from Tamil Nadu (14-17). Chikungunya which is endemic to South Asia, the Pacific island ar- ea, Africa, and the Americas and has infected millions of people mainly in developing coun- tries (18). The lack of a commercial vaccine and the failure of vector control strategies to limit the expansion of chikungunya have prompt- ed the need for further options to prevent the spread of this disease. Nowadays synthetic in- secticides are at the fore front of mosquito-con- trolling agents. The continual usage of the syn- thetic chemical insecticides possess various en- vironmental hazards such as development of resistance in vectors mosquitoes to these chem- icals, disruption of natural biological control systems in mosquito populations (19). Hence, the necessity of plant derived insecticides es- pecially target specific, eco-friendly, readily bi- odegradable and cost-effective (20). In general, plant essential oil has been recognized as im- portant natural resources of insecticides (21). Many researchers have reported the control of mosquito larvae using the plant extract and the essential oils obtained from the different parts of the plants (22-24). Natural insecticides meet the needs for alternatives to controlling resistant populations of different species of mosquitoes. They can affect different stages of development through a variety of mechanisms. In this study, we have chosen Elytraria acaulis belongs to the family Acanthaceae is a small shrub, which grows in shady dry places and it is commonly known as Asian Scalystem. It is a stem less perennial herb with one to several unbranched flowering stems; up to 30cm. Stems are cov- ered with overlapping bracts. Leaves occur in a rosette at the base. They are obovate, 4–10 centimeter long. Flowers are white, lower lip and lateral lobes spreading, 2-lobed. This plant is frequently found on rocky or sandy soils. The whole shrub is used for medicinal purposes (25). The decoction of E. acaulis leaves prescribed for fever, venereal diseases and root is used in mammary tumor, abscesses, pneumonia, anti- diabetic effects, antibacterial activity, treating wounds infected with worms and infantile di- arrhea as well as traditional medicine for long days (26-27). The sub-acute toxicity of meth- anolic extract of E. acaulis was tested against female Wistar rats with the concentrations of 50 to 2000mg/kg by oral administration (28). They observed that no significant alteration on any of the biological parameters. The present study was aimed to investigate the larvicidal properties of powdered leaf of E. acaulis against late third or early fourth instar larvae of Cx. quinquefasciatus and Ae. aegypti. Materials and Methods Selected medicinal plant Fresh and matured leaves of E. acaulis was used for the research work. The selected plant was collected from Kattukollai, Kanchipuram District, Tamil Nadu based on their abundance, availability, medicinal and insecticidal prop- erties. The plant specimen was identified by Dr P Paramasivam, Department of Botany, Pa- chaiyappa’s College for Men, Kanchipuram, India. The specimen plant was preserved at her- barium of Department of Botany, Pachaiyap- pa’s College for Men, Kanchipuram, India for further reference. The collected plant material were washed with tap water to remove all the unwanted impurities and shade dried at labor- atory temperature (27±2 °C) and macerated with electric blender and stored at 4 °C for larvicid- al bioassay. Maintenance of mosquito larvae Culex quinquefasciatus and Ae. aegypti mos- quito larvae were collected from stagnant wa- ter in and around of Kattukollai, Kanchipuram District, Tamil Nadu, India. All the larvae were kept in plastic trays containing tap water and were maintained in the laboratory. The larvae were fed with dog biscuits and yeast powder in the 3:1 ratio. All the experiments were car- ried out at 27±2 °C and 75–85% relative hu- http://jad.tums.ac.ir Published Online: September 30, 2020 http://jad.tums.ac.ir/ http://jad.tums.ac.ir/ J Arthropod-Borne Dis, September 2020, 14(3): 293–301 S Sukumaran and R Maheswaran: Larvicidal Activity of … 295 http://jad.tums.ac.ir Published Online: September 30, 2020 midity under a photoperiod of 14:10h (light/ dark) cycles. They were maintained until the larvicidal bioassay. Larvicidal activity Larvicidal activity was evaluated by using the standard method (29). Twenty five individ- uals of late third or early fourth instar larvae of Cx. quinquefasciatus and Ae. aegypti were re- leased in a 250ml glass beaker containing 100 ml of dechlorinated tap water mixed with de- sired E. acaulis plant powder at different con- centrations (mg), an equal number of controls were set up simultaneously using tap water. Five replicates of each concentration were run at a time. The experimental concentrations were 100, 120, 140, 160, 180 and 200mg/100ml respec- tively. Negative control (water) was run sim- ultaneously. Mortality and survival rate were recorded after 24 hours. Based on the WHO protocol no food was offered to avoid the dif- ference in mortality. The moribund and dead larvae in five replicates were combined and ex- pressed as a percentage of larval mortality for each concentration. Dead larvae were identi- fied when they failed to move after probing with a needle in the siphon or cervical region. Moribund larvae were those incapables of ris- ing to the surface (within reasonable period of time) or the water was disturbed the charac- teristic diving reaction was not seen. The LC50 value was calculated by EPA Probit analysis software. Results In the present investigation the toxic effect of powdered leaf of E. acaulis tested at six dif- ferent concentrations such as 200, 180, 160, 140, 120 and 100mg/100ml to evaluate the lar- vicidal activity against the larvae of Cx. Quin- quefasciatus and Ae. aegypti. Besides, the con- trol set up also compared with different con- centrations of plant powder. The highest con- centration (200mg/100ml) of powdered leaf of E. acaulis showed 100% mortality against the larvae of Cx. quinquefasciatus and Ae. aegypti. However, 180 and 160 and 140mg/100ml of E. acaulis leaf powder inflicted moderate larval mortality. The least concentrations of 120 and 100mg/100ml exhibited least larvicidal activi- ty. In comparison with the control, all the con- centrations of E. acaulis leaf powder contri- buted potential larvicidal activity. The LC50 and LC90 value of E. acaulis leaf powder ex- hibited 116.07 and 190.38mg/100ml against Cx. quinquefasciatus and 124.25 and 198.21 mg/100ml against Ae. aegypti (Table 1). No larval mortality of observed in control. After 96hrs 100% mortality was observed in all the tested concentrations E. acaulis. Symp- tomatological observations were carried out through the exposure period at laboratory tem- perature among the two species of mosquitoes revealed that immediately after exposure to E. acaulis. All larvae were active and exhibited a normal appearance with the siphon pointed up and head hung down. After 5 minutes of treat- ment, some of the larvae became restless and frequently sank down and floated up quickly at 200mg/100mlconcentration. At 30th minute, the restlessness persisted; tremor and convul- sion at the bottom of the container were ob- served approximately in 1 to 2 larvae. Similar evidences of restlessness, tremors, and con- vulsions followed by paralysis were clearly seen after an hour approximately in 4 to 5 larvae. At 12h, approximately 1 to 2 moribund and dead larvae were found. After 24h of treatment, ap- proximately one-third of the larvae was para- lyzed and sank to the bottom of the bowl. More and more larvae exhibited toxic symptoms dur- ing 12h. Subsequently, all of them died within 24 h in the 200mg/100ml treatment. The powdered leaf of E. acaulis caused rapid mortality, sug- gesting larvicidal property. The symptoms ob- served in treated larvae were similar to those caused by nerve poisons, such as excitation, convulsion, paralysis and death. Dead larvae were observed under the light microscope after 24h of exposure, where the body attained a dark brown color; length of the larvae was shrunk. http://jad.tums.ac.ir/ J Arthropod-Borne Dis, September 2020, 14(3): 293–301 S Sukumaran and R Maheswaran: Larvicidal Activity of … 296 http://jad.tums.ac.ir Published Online: September 30, 2020 Table 1. Larvicidal activity of Elytraria acaulis against the larvae of Culex quinquefasciatus and Aedes aegypti Culex quinquefasciatus Replication Concentration (mg/100ml)/ Number of dead out of 25 tested LC50 95% Confidence limit LC90 95% Confidence limit Control 200mg 180mg 160mg 140mg 120mg 100mg LCL UCL LCL UCL 1 0 25 20 19 15 13 10 116.07 90.05 131.08 190.38 164.26 279.40 2 0 25 20 20 15 13 9 3 0 25 21 20 16 13 9 4 0 25 20 20 18 14 11 5 0 25 20 19 16 12 11 Total dead 0/125 125/125 101/125 98/125 80/125 65/125 50/125 S.D. 0 0 0.44 0.54 1.22 0.70 1 % of mortality 0 100 80.8 78.4 64 52 40 Aedes aegypti 1 0 25 18 19 16 12 8 124.25 102.97 138.73 198.21 171.24 284.61 2 0 25 17 18 15 11 7 3 0 25 20 18 14 14 8 4 0 24 22 17 15 12 8 5 0 25 20 16 16 12 6 Total dead 0/125 124/125 97/125 88/125 76/125 61/125 37/125 S.D. 0 0.44 1.94 1.14 0.83 1.09 0.89 % of mortality 0 99.2 77.6 70.4 60.8 48.8 29.6 Values are mean ±SD of five replicates. In each concentration 25 larvae were used http://jad.tums.ac.ir/ J Arthropod-Borne Dis, September 2020, 14(3): 293–301 S Sukumaran and R Maheswaran: Larvicidal Activity of … 297 http://jad.tums.ac.ir Published Online: September 30, 2020 Discussions World Health Organization has estimated globally the contribution of commercial pesti- cides to health (30). Naturally occurring pes- ticides thus appear to have a prominent role in the development of future safety of the envi- ronment and public health (31). There are dif- ferent methods for controlling mosquitoes. The plant crude formulation was responsible for lar- vicidal activity. The pool of plants processing insecticidal substance is enormous. The use of plant essential extract for the pest and diseases management has recently been reversed. The preliminary screening is a good means of eval- uating the potential larvicidal activity of plants popularly used for this purpose. In the present investigation E. acaulis inflicted potential lar- vicidal activity with the LC50 value of 116.07 mg/100ml against Cx. quinquefasciatus and 124.25mg/100ml against Ae. aegypti. Like ways, the sustained toxicity test of some medicinal plants such as Nerium oleander, Calotropis procera and Ricinus communis powders against Anopheles arabiensis, Cx. quinquefasciatus (32). They reported that the after 6 days 100% mortality was observed in An. arabiensis, Whereas, Cx. quinquefasciatus 60% mortality was observed. The larvicidal activity due to the presence of Alkaloids, Flavonoids, Pro- tein, Amino Acid, Glycosides, Carbohydrates, Phenol, Steroids, Saponins and Tannins from E. acaulis (33). The results were coincides with earlier find- ings in which the leaf powder of Croton sparsiflorus had LC50 value of 122.73mg/100ml and LC90 value of 180.04mg/100ml followed by Bauhinia variegata with LC50 value of 142.47mg/100ml and LC90 value of 210.16mg/ 100ml, respectively (34). In another study the solvent extracts of E. acaulis showed mod- erate effect on Cx. quinquefasciatus and Ae. aegypti (35). Similarly, 1mg/ml of ethanolic extracts of the leaves of Lantana camara ex- hibited 84% larval mortality while treated with methanolic extract caused 48% mortality on fourth instar larvae of Ae. aegypti (36). The ethanolic extract from leaves of Cassia oc- cidentalis caused larval mortality against ma- larial vector mosquito An. stephensi at a dose equivalent to LC50 of 70.56% for fourth instar larvae (37). Ethanolic extract from bulbs of Allium sativum inflicted remarkable insecticidal activity against larvae of Aedes albopictus with LC50 value of 4.48g/L (38). The toxic effect of Ricinus communis crude extract was tested against immatures of Cx. Quinquefasciatus and An. arabiensis (39). They recorded LC50 values as 403.65, 445.66, and 498.88ppm against second, third, and fourth instar larvae of An. arabiensis and 1091.44, 1364.58, and 1445.44 ppm against second, third, and fourth instar larvae of Cx. quinquefasciatus, respectively Several plants were evaluated against main malaria vector, An. stephensi, and Cx. pipiens including Mentha spicata, Cymbopogon olivieri, Azadirachta indica, Melia azedarach, Lagetes minuta, Calotropis procera, Eucalyptus camal- dulensis, Cupressus arizonica, Thymus vul- garis, Lawsonia inermis, Cedrus deodara, Cionura erecta, Bunium persicum, Carum carvi, Artemisia dracunculus, Rosmarinus of- ficinalis. Mentha spicata and Eucalyptus camal- dulensis, had the lowest and highest LC50 respectively (40-55). Our results clearly indi- cated the E. acaulis highly effective against Cx. quinquefasciatus and Ae. aegypti. Powdered leaves of E. acaulis may be a good source to develop newer mosquitocidal biopesticide. Conclusion The overall findings of the present inves- tigation suggested that the E. acaulis highly effective against Cx. quinquefasciatus and Ae. aegypti larvae. However solvent extractions are time consuming and costlier technique. El- ytraria acaulis may be used as an alternative for synthetic chemical pesticides to control vec- http://jad.tums.ac.ir/ J Arthropod-Borne Dis, September 2020, 14(3): 293–301 S Sukumaran and R Maheswaran: Larvicidal Activity of … 298 http://jad.tums.ac.ir Published Online: September 30, 2020 tors mosquitoes and reduce vector borne diseases. According to our knowledge it seems not harmful to the natural environment and it needs more study to understand the level of toxicity. Acknowledgements The authors are thankful to Periyar Uni- versity, Salem, Tamil Nadu, India for the fi- nancial support. All authors declare that there is no conflict of interest. References 1. James AA (1992) Mosquito molecular ge- netics: the hands that feed bite back. Science. 257(5066): 37–38. 2. Peng Z, Yang J, Wang H, Simons FER (1999) Production and characterization of monoclonal antibodies to two new mosquito Aedes aegypti salivary proteins. Insect Biochem Mol Biol. 29: 909–914. 3. Bernhard L, Bernhard P, Magnussen P (2003) Management of patients with lymphoedema caused by filariasis in North-eastern Tanzania: alternative ap- proaches. Physiotheraphy. 89: 743–749. 4. Ramaiah KD, Eric AO (2014) Progress and impact of 13 years of the global pro- gramme to eliminate lymphatic filariasis on reducing the burden of filarial dis- ease. PLOS Negl Trop Dis. 8: e3319. 5. Richard HF, David RC (1959) Mosquitos of Medical Importance. U.S. Department of Agriculture, Washington D.C. 6. Simoy MI, Simoy MV, Canziani GA (2015) The effect of temperature on the popu- lation dynamics of Aedes aegypti. Ecol Model. 24: 100–110. 7. Kabra SK, Verma IC, Arora NK, Jain Y, Kalra V (1992) Dengue hemorrhagic fe- ver in children in Delhi. Bull World Health Org. 70(1): 105–108. 8. Agarwal R, Kapoor S, Nagar R, Misra A, Tandon R, Mathur A, Misra AK, Srivasta- va KL, Chaturvedi UC (1999) A clinical study of the patients with dengue he- morrhagic fever during the epidemic of 1996 at Lucknow, India. Southeast Asian J Trop Med Public Health. 30(4): 735– 740. 9. Kabra SK, Jain Y, Pandey RM, Madhulika, Singhal T, Tripathi P, Broor S, Seth P, Seth V (1999) Dengue hemorrhagic fever in children in the 1996 Delhi epidemic. Trans R Soc Trop Med Hyg. 93(3): 294– 298. 10. Dar L, Broor S, Sengupta S, Xess I, Seth P (1999) The first major outbreak of den- gue hemorrhagic fever in Delhi, India. Emerg Infect Dis. 5(4): 589–590. 11. Ram S, Khurana S, Kaushal V, Gupta R, Khurana SB (1998) Incidence of dengue fever in relation to climatic factors in Lu- dhiana, Punjab. Indian J Med Res. 108: 128–133. 12. Ilkal MA, Dhanda V, Hassan MM, Mava- le M, Mahadev PV, Shetty PS, Guttikar SN, Banerjee K (1991) Entomological in- vestigations during outbreaks of dengue fever in certain villages in Maharashtra state. Indian J Med Res. 93: 174–178. 13. Mahadev PV, Kollali VV, Rawal ML, Pu- jara PK, Shaikh BH, Ilkal MA, Pathak V, Dhanda V, Rodrigues FM, Banerjee K (1993) Dengue in Gujarat state, India during 1988 and 1989. Indian J Med Res. 97: 135–144. 14. Norman G, Theodre A, Joseph A (1991) An insular outbreak of dengue fever in a rural south Indian village. J Commun Dis. 23(3): 185–190. 15. Singh J, Balakrishnan N, Bhardwaj M, Amuthadevi P, George EG, Subramani K, Soundararajan K, Appavoo NC, Jain DC, Ichhpujani RL, Bhatia R, Sokhey J (2000) Silent spread of dengue and dengue hem- orrhagic fever to Coimbatore and Erode districts in Tamil Nadu, India, 1998: need for effective surveillance to monitor and http://jad.tums.ac.ir/ J Arthropod-Borne Dis, September 2020, 14(3): 293–301 S Sukumaran and R Maheswaran: Larvicidal Activity of … 299 http://jad.tums.ac.ir Published Online: September 30, 2020 control the disease. Epidemiol Infect. 125 (1): 195–200. 16. Cherian T, Ponnuraj E, Kuruvilla T, Kiruba- karan C, John TJ, Raghupathy P (1994) An epidemic of dengue hemorrhagic fever and dengue shock syndrome in and around Vellore. Indian J Med Res. 100: 51–56. 17. Abdul KM, Kandaswamy P, Appavoo NC (1997) Outbreak and control of dengue in a village in Dharmapuri, Tamil Nadu. J Commun Dis. 29(1): 69–71. 18. Vega-Rúa A, Lourenço-de-Oliveira R, Mous- son L, Vazeille M, Fuchs S, Yébakima A, Gustave J, Girod R, Dusfour I, Leparc- Goffart I, Vanlandingham DL, Huang YJS, Lounibos LP, Ali SM, Nougairede A, Lamballerie X, Failloux AB (2015) Chikungunya virus transmission potential by local Aedes mosquitoes in the Americas and Europe. PLoS Negl Trop Dis. 9 (5): e0003780. 19. Pushpanathan T, Jebanesan A, Govindara- jan M (2008) The essential oil of Zingi- ber officinalis Linn. (Zingiberaceae) as a mosquito larvicidal and repellent agent against the filarial vector Culex quinque- fasciatus Say (Diptera: Culicidae). Para- sitol Res. 102(6): 1289–1291. 20. Rajkumar S, Jebanesan A (2005) Oviposi- tion deterrent and skin repellent activi- ties of Solanum trilobatum leaf extract against the malarial vector Anopheles stephensi. J Insect Sci. 5: 15. 21. Maheswaran R, Ignacimuthu S (2015) Ef- fect of confertifolin from Polygonum hy- dropiper L. against dengue vector mos- quitoes Aedes aegypti L. Environ Sci Pollut Res Int. 22(11): 8280–8287. 22. Govindarajan M, Rajeswary M (2015) Ovi- cidal and adulticidal potential of leaf and seed extract of Albizia lebbeck (L.) Benth. (Family: Fabaceae) against Culex quin- quefasciatus, Aedes aegypti, and Anopheles stephensi (Diptera: Culici-dae). Parasitol Res. 114(5): 1949–1961. 23. Maheswaran R, Sathish S, Ignacimuthu S (2008) Larvicidal activity of Leucus aspera (Willd.) against the larvae of Culex quin- quefasciatus Say and Aedes aegypti L. Int J Integ Biol. 2(3): 214–217. 24. Benelli G (2015) Research in mosquito con- trol: current challenges for a brighter future. Parasitol Res. 114(8): 2801–2805. 25. Ravikanth V, Ramesh P, Diwan PV, Ven- kateswarlu Y (2001) Pyrazole alkaloids from Elytraria acaulis. Biochem Syst Ecol. 29(7): 753–754. 26. Krishna GRD (2013) Hypoglycemic activ- ity of extracts from Elytraria acaulis L. leaves in alloxan- induced diabetic rats. Biolife, 1: 11–16. 27. Kowsalya V (2012) Antibacterial activity of honey and Elytraria acaulis against bacteria isolated from burnt wound sep- sis. J Pharm Biol Sci. 1(5): 1–20. 28. Koshy RK, Kapoor BR, Azmathulla M (2011) Acute and subacute toxicity of methanol extract of Elytraria acaulis lan- dau in rat. Pharmacology online. 3: 229– 242. 29. Patil PB, Holihosur SN, Kallapur VL (2006) Efficacy of natural product, Clerodendron inerme against dengue mosquito vector Aedes aegypti. Curr Sci. 90(8): 1064– 1066. 30. World Health Organization (1992) Lym- phatic filariasis: the disease and its con- trol. Fifth report of the expert committee on filariasis. WHO Technical Report Se- ries. 821: 1–71. 31. Mandava NB (1985) The chemistry of al- lopathy, biochemical interactions among plants. Thomson AC (Ed) ACS sympo- sium series 268, Am Chem Soc Wash- ington, pp. 33–54. 32. Kehail MAA, Bashir NHH, Abdelrahman EE, Abdelrahim AM (2017) Larvicidal activity of three plants powders and aque- ous extracts on Anopheles and Culex mosquito larvae (Diptera: Culicidae). Int J Mosq Res. 4(4): 37–41. 33. Kiruthika N, Dhivya R, Kalaiselvi K, Kan http://jad.tums.ac.ir/ J Arthropod-Borne Dis, September 2020, 14(3): 293–301 S Sukumaran and R Maheswaran: Larvicidal Activity of … 300 http://jad.tums.ac.ir Published Online: September 30, 2020 imozhi P, Panneerselvam K (2012) Phy- tochemical studies on Elytraria acaulis. Int J Pharm Bio Sci. 3(3): 1054–1062. 34. Shanmugapriya R, Maheswaran R, Igna- cimuthu S (2015) Bauhinia variegata L. and Croton sparsiflorus L. against the lar- vae of Aedes aegypti L. J Zool Sci. 3(2): 1–4. 35. Rajiv Gandhi M, Daniel Reegan A, Sivasan- karan K, Gabriel Paulraj M, Ignacimuthu S (2016) Ovicidal and larvicidal activi- ties of some plant extracts against Aedes aegypti L. and Culex quinquefasciatus Say (Diptera: Culicidae). Asian Pac J. Trop Dis. 6(6): 468–471. 36. Kumar MS, Maneemegalai S (2008) Eva- luation of larvicidal effect of Lantana camara Linn. against mosquito species Ae- des aegypti and Culex quinquefasciatus. Adv Biol Res. 2: 39–43. 37. Dhandapani A, Kadarkarai M (2011) HPTLC quantification of flavonoids, lar- vicidal and smoke repellent activities of Cassia occidentalis L. (Caesalpiniaceae) against malarial vector Anopheles stephen- si Liston (Diptera: Culicidae). J Phytol. 3: 60–72. 38. Tedeschi P, Leis M, Pezzi M, Civolani S, Maietti A, Brandolini V (2011) Insecti- cidal activity and fungitoxicity of plant extracts and components of horseradish (Armoracia rusticana) and garlic (Allium sativum). J Environ Sci Health B. 46: 486–490. 39. Elimam AM, Elmalik KH, Ali FS (2009) Larvicidal, adult emergence inhibition and oviposition deterrent effects of foliage extract from Ricinus communis L. against Anopheles arabiensis and Culex quinquefasciatus in Sudan. Trop Biomed. 26: 130–139. 40. Hajiakjoondi A, Aghel N, Xamanizadeh- Nadgar N, Vatandoost H (2008) Chemi- cal and biological study of Mentha spi- cata L. essential oil from Iran. Daru. 8 (1–2): 19–21. 41. Hadjiakhoondi A, Vatandoost H, Jamshidi AH, Bagherj Amiri E (2003) Chemical constituents and efficacy of Cymbopogon olivieri (Boiss) bar essential oil against malaria vector, Anopheles stephensi. Daru. 11(3): 125–128. 42. Vatandoost H, MoinVaziri V (2004) Lar- vicidal activity of a neem tree extract (Neemarin) against mosquito larvae in the Islamic Republic of Iran. East Medi- terr Health J. 10(4/5): 573–581. 43. Hadjiakhoondi A, Vatandoost H, Khanavi M, Sadeghipour-Roodsari HR, Vosoughi M, Kazemi M, Abai MR (2006) Fatty acid composition and toxicity of Melia azedarach L. Fruits against Malaria Vec- tor Anopheles stephensi. Iran J Pharma- ceutical Sci. 2(2): 97–102. 44. Hajiakhondi A, Vatandoost H, Abousaber H, Khanavai M, Abdi L (2008) Chemi- cal composition of the essential oil of Tagetes minuta L and its effect on Anoph- eles stephensi larvae in Iran. J Med Plants. 7(26): 33–39. 45. Shahi M, Hanafi-Bojd AA, Iranshahi M, Vatandoost H, Hanafi-Bojd MY (2010) Larvicidal efficacy of latex and extract of Calotropis procera (Gentianales: Ascle- piadaceae) against Culex quinquefasciatus and Anopheles stephensi (Diptera: Cu- licidae). J Vector Borne Dis. 47(3): 185– 188. 46. Sedaghat MM, Sanei-Dehkhordi AR, Kha- navi M, Abai MR, Hadjiiakhondi A, Moh- tarami F, Vatandoost H (2010) Phyto- chemistry and larvicidal activity of Eu- calyptus camaldulensis against malaria vector, Anopheles stephensi. Asian Pacific J Trop Med. 3(11): 841–845. 47. Sedaghat MM, Sanei-Dehkordi AR, Kha- navi M, Abai MR, Mohtarami F, Vatan- doost H (20110) Chemical composition and larvicidal activity of essential oil of Cupressus arizona E.L. Greene against malaria vector Anopheles stephensi Lis- ton (Diptera: Culicidae). Pharmacognosy http://jad.tums.ac.ir/ http://www.openj-gate.org/Search/SearchResults.aspx?SearchTerm=%22H.%20Vatandoost%22&Field=AU&res=10&type=2&sub=All&update=none&from=-1&to=2011&pr=2 http://www.openj-gate.org/Search/SearchResults.aspx?SearchTerm=%22V.M.%20Vaziri%22&Field=AU&res=10&type=2&sub=All&update=none&from=-1&to=2011&pr=2 http://www.ncbi.nlm.nih.gov/pubmed/20834091 http://www.ncbi.nlm.nih.gov/pubmed/20834091 http://www.ncbi.nlm.nih.gov/pubmed/20834091 http://www.ncbi.nlm.nih.gov/pubmed/20834091 http://www.ncbi.nlm.nih.gov/pubmed/20834091 J Arthropod-Borne Dis, September 2020, 14(3): 293–301 S Sukumaran and R Maheswaran: Larvicidal Activity of … 301 http://jad.tums.ac.ir Published Online: September 30, 2020 Res. 3(2): 135–139. 48. Khanavi M, Rajabi A, Behzad M, Hadji- akhoondi A, Vatandoost H, Abai MR (2011) Larvicidal activity of Centaurea bruguierana ssp. belangerana against Anopheles stephensi. Iran J Pharm Res. 10(4): 829–833. 49. Khanavi M, Alireza F, Vatandoost H, Sed- aghat MM, Abai MR, Hadjiakhoondi A (2012) Larvicidal activity of essential oil and methanol extract of Nepeta men- thoides against malaria vector, Anophe- les stephensi. Asian Pac J Trop Med. 5 (12): 962–965. 50. Vatandoost H, Sanei-Dehkordi A, Sadeghi SM, Davari B, Karimian F, Abai MR, Sedaghat AA (2012) Identification of chemical constituents and larvicidal ac- tivity of Kelussia odoratissima Mozaf- farian essential oil against two mosquito vectors Anopheles stephensi and Culex pipiens (Diptera: Culicidae). Experimental Parasitol. 132(4): 470–474. 51. Khanavi M, Vatandoost H, Dehaghi NK, Sanei-Dehkordi A, Sedaghat MM, Hadjiakhoondi A, Hadjiakhoondi F (2013) Larvicidal activity of some Iranian plants against malaria vector An. stephensi. Acta Medica Iranica. 51(3): 141–147. 52. Mozaffari E, Abai MR, Khanavi M, Vatan- doost H, Sedaghat MM, Sanei-Dehkordi A, Rafi F (2014) Chemical composition, larvicidal and repellent properties of Cionura erecta (L.) Griseb. against ma- laria vector, Anopheles stephensi Liston (Diptera: Culicidae) under laboratory con- ditions. J Arthropod Borne Dis. 8(2): 147– 155. 53. Golfakhrabadi F, Khanavi M, Ostad SN, Saeidnia S, Vatandoost H, Abai MR, Hafizi M, Yousefbeyk F, Razzaghi-Rad Y, Ameneh Baghenegadian A, Shams- Ardekani MR (2015) Biological activi- ties and composition of Ferulago cardu- chorum essential oil. J Arthropod Borne Dis. 9(1): 104–115. 54. Sanei-Dehkordi A, Vatandoost H, Abai MR, Davari B, Sedaghat M (2016) Chemical composition and larvicidal activity of Bunium persicum essential oil against two important mosquito vectors. J Essent Oil-Bear Plants. 19(2): 49–357. 55. Torabi Pour H, Shayeghi M, Vatandoost H, Abai MR (2016) Study on larvicidal effects of essential oils of three Iranian native plants against larvae of Anopheles stephensi (Liston). Vector Biol J. 1(2): 2–6. http://jad.tums.ac.ir/ https://www.ncbi.nlm.nih.gov/pubmed/?term=Hafizi%20M%5BAuthor%5D&cauthor=true&cauthor_uid=26114148 https://www.ncbi.nlm.nih.gov/pubmed/?term=Yousefbeyk%20F%5BAuthor%5D&cauthor=true&cauthor_uid=26114148 https://www.ncbi.nlm.nih.gov/pubmed/?term=Rad%20YR%5BAuthor%5D&cauthor=true&cauthor_uid=26114148 https://www.ncbi.nlm.nih.gov/pubmed/?term=Baghenegadian%20A%5BAuthor%5D&cauthor=true&cauthor_uid=26114148 https://www.ncbi.nlm.nih.gov/pubmed/?term=Ardekani%20MR%5BAuthor%5D&cauthor=true&cauthor_uid=26114148 https://www.ncbi.nlm.nih.gov/pubmed/?term=Ardekani%20MR%5BAuthor%5D&cauthor=true&cauthor_uid=26114148